Therapeutic agent for dyslipidemia

ABSTRACT

The present invention addresses the problem of providing a pharmaceutical combination composition and drug combinations for the prevention and/or treatment of dyslipidemic conditions such as atherosclerosis, hypercholesterolemia, low HDL blood disease in mammals including humans. A pharmaceutical composition for the prevention and/or treatment of dyslipidemia, etc. which comprises (a) a compound represented by the general formula (1) (in the formula, each symbol is the same as defined in the specification) or a salt thereof or a solvate thereof and (b) a cholesterol absorption inhibitor.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.15/399,856, filed on Jan. 6, 2017, which is a continuation of U.S.application Ser. No. 14/429,449, filed on Mar. 19, 2015, which is a 371of International Application No. PCT/JP2013/005756, filed on Sep. 27,2013, which claims the benefit of priority from the prior JapanesePatent Application No. 2012-214635, filed on Sep. 27, 2012, the entirecontents of which are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to a composition containing a compound (1)selectively activating an α-type (PPARα) of peroxisomeproliferator-activated receptors (PPARs), and a cholesterol absorptioninhibitor, which are intended to prevent and/or treat atherosclerosis,and a lipemia condition such as hypercholesterolemia; and a combinationthereof.

BACKGROUND ART

In recent years, due to the westernization of diet, patients withhypercholesterolemia, hypertriglyceridemia, hypo-HDL chloesterolemia,and the like, which are in the category of a so-called lifestyle-relateddiseases, are in an increasing trend. Further, recently, patients withmixed or combined dyslipidemia, which has both hypercholesterolemia andhypertriglyceridemia, have been increased. Especially in patients withmixed dyslipidemia, LDL cholesterol (LDL-C) and triglyceride (TG) areincreased, and HDL cholesterol (HDL-C) is decreased. Such a high TG andlow HDL-C condition is also observed in patients with metabolic syndromeor diabetes. It has been proved that hyper LDL-cholesterolemia, hypo-HDLcholesterolemia, and hypertriglyceridemia are risk factors for coronaryartery disease (CAD) or cerebral vascular disorder and the like. Theimportance of the management of dyslipidemia is described in “Guidelinesfor Preventing Arteriosclerotic Diseases, 2012 edition” released byJapan Atherosclerosis Society.

Dyslipidemia, in particular, hypercholesterolemia has already falleninto a disease area of high medical satisfaction with the advent ofstatins. However, from the results of a large number of large-scaleclinical trials, it has been found that the further decrease of bloodLDL cholesterol level leads to the prevention of coronary artery disease(the lower the better), and the more strict lipid control isrecommended. There are a large number of patients who cannot reach theintended level of blood LDL-C only by the statins, and treatment with acombination of multiple agents has been required. On the other hand, forthe hypertriglyceridemia, it has been indicated that fibrate-basedagents more effectively reduce the level, and for the hypo-HDLcholesterolemia, research and development of CETP inhibitors, apo A-Iincreasing agents, and the like has been carried out. In particular, ithas been expected that blood HDL-C increasing drugs are increasinglyimportant in the future from the viewpoint of reverse transfer ofcholesterol, and evolution suppression and regression ofarteriosclerosis, (Non-Patent Document 1).

PPAR is one of the receptors that belong to a nuclear receptor family.It has been known of the presence of three subtypes (α, γ, and δ) in thereceptor (Non-Patent Document 2). Among them, PPARα is mainly expressedin the liver, and when PPARα is activated, production of apo C-III issuppressed, and then lipoprotein lipase (LPL) is activated, as a result,fat is decomposed. As the PPARα agonist, unsaturated fatty acids;fibrate-based agents such as fenofibrate, bezafibrate, and gemfibrozil;and the like have been known (Non-Patent Document 3). Further, in recentyears, a compound, which has a PPARα activating effect more strongly andselectively than the conventional fibrate-based agent does, has beenreported (Patent Documents 1 to 10).

A cholesterol absorption inhibitor is an agent suppressing theabsorption of dietary cholesterol from the small intestine. As to thefactors to determine the blood cholesterol level, it is considered thatcontribution of de novo synthesized cholesterol in the liver is 70%, andcontribution of the dietary cholesterol is 30%, therefore, the bloodcholesterol levels can be reduced by the suppression of the absorptionof dietary cholesterol from the small intestine, independently from thecholesterol synthesis inhibition by statins. As the cholesterolabsorption inhibitor, for example, ezetimibe (Zetia (registeredtrademark)) has been known. The mechanism of its action was unknown fora long time, however, recently the mechanism that the absorption ofcholesterol is suppressed by the inhibition of a cholesterol transporter(Niemann-Pick C1-like 1, NPC1-L1) expressed in the small intestinalepithelial cells has been elucidated. In addition to ezetimibe, multiplecompounds having an inhibitory effect on the cholesterol transporterhave been reported (Patent Documents 11 to 14).

Treatments with a combination of the above-described fibrate-based agentwith ezetimibe have been investigated for patients with mixeddyslipidemia who have a feature that the blood LDL-C level and the bloodTG level are increased, and the blood HDL-C level is decreased (PatentDocument 15, and Non-Patent Documents 4 and 5). Meanwhile, it has notbeen known what kind of effect is exerted on the dyslipidemia if acompound (1) of the present invention is used in combination withezetimibe.

CITATION LIST Patent Document

-   Patent Document 1: WO 2005/023777-   Patent Document 2: WO 2009/080248-   Patent Document 3: WO 2009/047240-   Patent Document 4: WO 2008/006043-   Patent Document 5: WO 2006/049232-   Patent Document 6: WO 2006/033891-   Patent Document 7: WO 2005/009942-   Patent Document 8: WO 2004/103997-   Patent Document 9: WO 2005/097784-   Patent Document 10: WO 2003/043997-   Patent Document 11: WO 2007/008541-   Patent Document 12: WO 2007/008529-   Patent Document 13: WO 2008/033464-   Patent Document 14: WO 2008/104875-   Patent Document 15: WO 2002/058732

Non-Patent Document

-   Non-Patent Document 1: Folia Pharmacol. Jpn., 129, 267-270 (2007)-   Non-Patent Document 2: J. Lipid Research 37, 907-925 (1996)-   Non Patent Document 3: Trends in Endocrinology and Metabolism,    15(7), 324-330 (2004)-   Non-Patent Document 4: European Heart Journal, 26, 897-905 (2005)-   Non-Patent Document 5: Clinical Medicine: Therapeutics, 1, 1703-1713    (2009)

SUMMARY OF THE INVENTION Problems to be Solved in the Invention

An object of the present invention is to provide a pharmaceuticalcombination composition and a drug combination for prevention and/ortreatment of atherosclerosis, or a dyslipidemic condition such ashypercholesterolemia, and hypo-HDL cholesterolemia, and the like.

Means for Solving the Problems

As a result of intensive study, in view of the above circumstances, thepresent inventors found the fact that a strong blood HDL cholesterolincreasing effect is exerted by a combination of a phenoxyacetic acidderivative represented by the following general formula (1) or a saltthereof, with ezetimibe that is a cholesterol absorption inhibitor,wherein the phenoxyacetic acid derivative represented by the followinggeneral formula (1) is disclosed in the Patent Document 1 to have aselective PPARα activating effect, not to be accompanied by weight gainor obesity in mammals including human beings, and to be useful as aprophylactic and/or therapeutic agent for dyslipidemia,arteriosclerosis, diabetes, diabetic complication (diabetic nephropathyand the like), inflammation, heart disease, and the like, and thus havecompleted the present invention.

That is, the present invention is to provide a pharmaceuticalcomposition for preventing and/or treating dyslipidemia, including:

-   a) a compound represented by the following general formula (1):

-   wherein R¹ and R², which may be identical or different, each    represents a hydrogen atom, a methyl group, or an ethyl group;    R^(3a), R^(3b), R^(4a) and R^(4b), which may be identical or    different, each represents a hydrogen atom, a halogen atom, a nitro    group, a hydroxyl group, a C₁₋₄ alkyl group, a trifluoromethyl    group, a C₁₋₄ alkoxy group, a C₁₋₄ alkylcarbonyloxy group, a di-C₁₋₄    alkylamino group, a C₁₋₄ alkylsulfonyloxy group, a C₁₋₄    alkylsulfonyl group, a C₁₋₄ alkylsulfinyl group, or a C₁₋₄ alkylthio    group, or R^(3a) and R^(3b), or R^(4a) and R^(4b) are joined to    represent an alkylenedioxy group; X represents an oxygen atom, a    sulfur atom, or N—R⁵ (wherein R⁵ represents a hydrogen atom, a C₁₋₄    alkyl group, a C₁₋₄ alkylsulfonyl group, or a C₁₋₄ alkyloxycarbonyl    group); Y represents an oxygen atom, a S(O)₁ group (wherein 1    represents 0, 1 or 2), a carbonyl group, a carbonylamino group, an    aminocarbonyl group, a sulfonylamino group, an aminosulfonyl group,    or an NH group; Z represents CH, or N; n represents a number from 1    to 6; and m represents a number from 2 to 6, or a salt thereof, or a    solvate of the compound, or a solvate of the salt of the compound;    and-   b) a cholesterol absorption inhibitor.

In more detail, the present invention is to provide a pharmaceuticalcomposition for preventing and/or treating mixed dyslipidemia, metabolicsyndrome, dyslipidemia complicated by diabetes, hypo-HDLcholesterolemia, or hypertriglyceridemia, including: a) a compoundrepresented by the general formula (1) or a salt thereof, or a solvateof the compound, or a solvate of the salt of the compound; and b) acholesterol absorption inhibitor.

The more detailed description of the present invention is as follows.

-   (1) A pharmaceutical composition for preventing and/or treating    dyslipidemia, including: a compound represented by the general    formula (1) or a salt thereof, or a solvate of the compound, or a    solvate of the salt of the compound; and a cholesterol absorption    inhibitor.-   (2) The pharmaceutical composition according to (1), wherein the    compound represented by the general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (3) The pharmaceutical composition according to (1) or (2), wherein    the cholesterol absorption inhibitor is ezetimibe.-   (4) The pharmaceutical composition according to any one of (1) to    (3), wherein the dyslipidemia is a disease selected from the group    consisting of mixed dyslipidemia, metabolic syndrome, dyslipidemia    complicated by diabetes, hypo-HDL chloesterolemia (hypo HDL-C-emia),    and hypertriglyceridemia (hyper TG-emia).-   (5) The pharmaceutical composition according to any one of (1) to    (4), wherein an amount ratio of the compound represented by the    general formula (1) to the cholesterol absorption inhibitor is 1:200    to 3:1.-   (6) A pharmaceutical composition for increasing HDL cholesterol    (HDL-C), including: a compound represented by the general    formula (1) or a salt thereof, or a solvate of the compound, or a    solvate of the salt of the compound; and a cholesterol absorption    inhibitor.-   (7) The pharmaceutical composition according to (6), wherein the    compound represented by the general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (8) The pharmaceutical composition according to (6) or (7), wherein    the cholesterol absorption inhibitor is ezetimibe.-   (9) The pharmaceutical composition according to any one of (6) to    (8), wherein a disease requiring increase of HDL cholesterol (HDL-C)    is a disease selected from the group consisting of mixed    dyslipidemia, metabolic syndrome, dyslipidemia complicated by    diabetes, hypo-HDL chloesterolemia (hypo HDL-C-emia), and    hypertriglyceridemia (hyper TG-emia).-   (10) The pharmaceutical composition according to any one of (6) to    (9), wherein an amount ratio of the compound represented by the    general formula (1) to the cholesterol absorption inhibitor is 1:200    to 3:1.-   (11) A method for preventing and/or treating dyslipidemia in a    subject, comprising: administering to a subject with dyslipidemia or    a subject with a risk of dyslipidemia an effective amount of a    pharmaceutical composition containing a compound represented by the    general formula (1) or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound, and a cholesterol    absorption inhibitor.-   (12) The method for preventing and/or treating dyslipidemia in a    subject according to (11), wherein the compound represented by the    general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (13) The method for preventing and/or treating dyslipidemia in a    subject according to (11) or (12), wherein the cholesterol    absorption inhibitor is ezetimibe.-   (14) The method for preventing and/or treating dyslipidemia with a    subject according to any one of (11) to (13), wherein the    dyslipidemia is selected from the group consisting of mixed    dyslipidemia, metabolic syndrome, dyslipidemia complicated by    diabetes, hypo-HDL chloesterolemia (hypo HDL-C-emia), and    hypertriglyceridemia (hyper TG-emia).-   (15) The method for preventing and/or treating dyslipidemia in a    subject according to anyone of (11) to (14), wherein an amount ratio    of the compound represented by the general formula (1) to the    cholesterol absorption inhibitor in the pharmaceutical composition    is 1:200 to 3:1.-   (16) A method for increasing HDL cholesterol (HDL-C) in a subject,    comprising: administering to a subject requiring increase of HDL    cholesterol (HDL-C) an effective amount of a pharmaceutical    composition containing a compound represented by the general    formula (1) or a salt thereof, or a solvate of the compound, or a    solvate of the salt of the compound, and a cholesterol absorption    inhibitor.-   (17) The method according to (16), wherein the compound represented    by the general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (18) The method according to (16) or (17), wherein the cholesterol    absorption inhibitor is ezetimibe.-   (19) The method according to any one of (16) to (18), wherein a    disease in a subject requiring increase of HDL cholesterol (HDL-C)    is selected from the group consisting of mixed dyslipidemia,    metabolic syndrome, dyslipidemia complicated by diabetes, hypo-HDL    chloesterolemia (hypo HDL-C-emia), and hypertriglyceridemia (hyper    TG-emia).-   (20) The method according to any one of (16) to (19), wherein an    amount ratio of the compound represented by the general formula (1)    to the cholesterol absorption inhibitor in the pharmaceutical    composition is 1:200 to 3:1.-   (21) A compound represented by the general formula (1) or a salt    thereof, or a solvate of the compound, or a solvate of the salt of    the compound, for use as a pharmaceutical combination composition    for preventing and/or treating dyslipidemia in combination with a    cholesterol absorption inhibitor.-   (22) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to-   (21), wherein the compound represented by the general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (23) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to (21)    or (22), wherein the cholesterol absorption inhibitor is ezetimibe.-   (24) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to any    one of (21) to (23), wherein the dyslipidemia is selected from the    group consisting of mixed dyslipidemia, metabolic syndrome,    dyslipidemia complicated by diabetes, hypo-HDL chloesterolemia (hypo    HDL-C-emia), and hypertriglyceridemia (hyper TG-emia).-   (25) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to any    one of (21) to (24), wherein an amount ratio of the compound    represented by the general formula (1) to the cholesterol absorption    inhibitor in the pharmaceutical composition is 1:200 to 3:1.-   (26) A compound represented by the general formula (1) or a salt    thereof, or a solvate of the compound, or a solvate of the salt of    the compound, for use as a pharmaceutical combination composition    for increasing HDL cholesterol (HDL-C) in combination with a    cholesterol absorption inhibitor.-   (27) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to (26),    wherein the compound represented by the general formula (1) is    (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric    acid.-   (28) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to (26)    or (27), wherein the cholesterol absorption inhibitor is ezetimibe.-   (29) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to any    one of (26) to (28), wherein a disease in a subject requiring    increase of HDL cholesterol (HDL-C) is selected from the group    consisting of mixed dyslipidemia, metabolic syndrome, dyslipidemia    complicated by diabetes, hypo-HDL chloesterolemia (hypo HDL-C-emia),    and hypertriglyceridemia (hyper TG-emia).-   (30) The compound or a salt thereof, or a solvate of the compound,    or a solvate of the salt of the compound for use according to anyone    of (26) to (29), wherein an amount ratio of the compound represented    by the general formula (1) to the cholesterol absorption inhibitor    in the pharmaceutical composition is 1:200 to 3:1.

Effects of the Invention

The agent and pharmaceutical composition of the present invention havean excellent blood HDL cholesterol increasing effect, and are useful inthe prevention and/or treatment of dyslipidemia, particularly mixeddyslipidemia, metabolic syndrome, dyslipidemia complicated by diabetes,hypo-HDL cholesterolemia, or hypertriglyceridemia.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 illustrates plasma HDL-C levels when compound A and ezetimibe areadministered each alone or in combination.

FIG. 2 illustrates plasma TC levels when compound A and ezetimibe areadministered each alone or in combination.

FIG. 3 illustrates plasma VLDL-C levels when compound A and ezetimibeare administered each alone or in combination.

FIG. 4 illustrates plasma LDL-C levels when compound A and ezetimibe areadministered each alone or in combination.

FIG. 5 illustrates plasma HDL-C levels when compound A and ezetimibe areadministered each alone or in combination.

FIG. 6 illustrates plasma HDL-C levels when fenofibrate and ezetimibeare administered each alone or in combination.

MODES FOR CARRYING OUT THE INVENTION

Examples of the halogen atom of R^(3a), R^(3b), R^(4a), and R^(4b) inthe general formula (1) include a fluorine atom, a chlorine atom, and abromine atom. Among them, a fluorine atom and a chlorine atom areparticularly preferable.

Examples of the C₁₋₄ alkyl group of R^(3a), R^(3b), R^(4a), R^(4b), andR⁵ include a methyl group, an ethyl group, an n-propyl group, anisopropyl group, and a butyl group. Among them, a methyl group isparticularly preferable.

Examples of the C₁₋₄ alkoxy group of R^(3a), R^(3b), R^(4a) and R^(4b)include a methoxy group, an ethoxy group, an n-propoxy group, anisopropoxy group, and a butoxy group. Among them, a methoxy group isparticularly preferable.

Examples of the C₁₋₄ alkylcarbonyloxy group of R^(3a), R^(3b), R^(4a)and R^(4b) include a methylcarbonyloxy group, an ethylcarbonyloxy group,an n-propylcarbonyloxy group, an isopropylcarbonyloxy group, and abutylcarbonyloxy group. Among them, a methylcarbonyloxy group isparticularly preferable.

Examples of the di-C₁₋₄ alkylamino group of R^(3a), R^(3b), R^(4a) andR^(4b) include a dimethylamino group, a diethylamino group, and adiisopropylamino group. Among them, a dimethylamino group isparticularly preferable.

Examples of the C₁₋₄ alkylsulfonyloxy group of R^(3a), R^(3b), R^(4a)and R^(4b) include a methylsulfonyloxy group, and an ethylsulfonyloxygroup. Among them, a methylsulfonyloxy group is particularly preferable.

Examples of the C₁₋₄ alkylsulfonyl group of R^(3a), R^(3b), R^(4a),R^(4b), and R⁵ include a methylsulfonyl group, and an ethylsulfonylgroup. Among them, a methylsulfonyl group is particularly preferable.

Examples of the C₁₋₄ alkylsulfinyl group of R^(3a), R^(3b), R^(4a) andR^(4b) include a methylsulfinyl group, and an ethylsulfinyl group. Amongthem, a methylsulfinyl group is particularly preferable.

Examples of the C₁₋₄ alkylthio group of R^(3a), R^(3b), R^(4a) andR^(4b) include a methylthio group, and an ethylthio group. Among them, amethylthio group is particularly preferable.

Examples of the alkylenedioxy group in which R^(3a) and R^(3b), orR^(4a) and R^(4b) are bonded to each other include a methylenedioxygroup, and an ethylenedioxy group. Among them, a methylenedioxy group isparticularly preferable.

Examples of the C₁₋₄alkyloxycarbonyl group of R⁵ include amethyloxycarbonyl group, and an ethyloxycarbonyl group. Among them, amethyloxycarbonyl group is particularly preferable.

R¹ and R² are particularly preferably hydrogen atoms at the same time,or methyl groups at the same time; or a methyl group and a hydrogenatom, or an ethyl group and a hydrogen atom.

X represents an oxygen atom, a sulfur atom, or N—R⁵, and is preferablyan oxygen atom. Further, Y represents an oxygen atom, a S(O)₁ group, acarbonyl group, a carbonylamino group, an aminocarbonyl group, asulfonylamino group, an aminosulfonyl group, or an NH group, and ispreferably an oxygen atom. Z represents CH or N, and is preferably CH. 1represents 0, 1 or 2, and is preferably 2. n represents a number from 1to 6, and is preferably from 1 to 3. m represents a number from 2 to 6,is preferably 2 to 4, and is particularly preferably 2 or 3.

Preferable examples of the compound represented by the general formula(1) of the present invention include a compound in which X and Y areoxygen atoms, Z is CH, n is 1, and m is 3. More preferable examples ofthe compound include a compound in which X and Y are oxygen atoms; Z isCH; n is 1; m is 3; R^(3a) and R^(3b), which may be identical ordifferent, each of which is a hydrogen atom, a halogen atom, a hydroxylgroup, a C₁₋₄ alkyl group, a trifluoromethyl group, or a C₁₋₄ alkoxygroup; R^(4a) and R^(4b), which may be identical or different, each ofwhich is a halogen atom, a hydroxyl group, a C₁₋₄ alkyl group, atrifluoromethyl group, or a C₁₋₄ alkoxy group. More preferable examplesof the compound represented by the general formula (1) of the presentinvention include(R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyricacid (hereinafter, may be referred to as “compound A”).

Usually, in an amount of around 400 to 500 mg of cholesterol from thediet, and in an amount of around 800 to 2,000 mg of cholesterol from thebile are drawn into the small intestine per day, and around 50% of thecholesterol drawn into the small intestine is absorbed. In the presentinvention, the cholesterol absorption inhibitor is not particularlylimited as long as being able to inhibit the absorption of thecholesterol drawn into the small intestine, and examples of thecholesterol absorption inhibitor include, for example, an anion exchangeresin such as cholestyramine, and cholestyramide, and an agentsuppressing the absorption of cholesterol by the inhibition of acholesterol transporter (NPC1-L1), such as ezetimibe, and a compounddescribed in the Patent Documents 11 to 14. Preferable examples of thecholesterol absorption inhibitor of the present invention include, forexample, an agent suppressing the absorption of cholesterol by theinhibition of a cholesterol transporter (NPC1-L1), and among them,ezetimibe is more preferable.

A compound represented by the general formula (1) of the presentinvention can be produced, for example, in accordance with a methoddescribed in Patent Document 1.

In addition, in the present invention, a salt or solvate of the compoundrepresented by the general formula (1) can be used. The salt and solvatecan be produced by routine procedures.

The salt of the compound represented by the general formula (1) of thepresent invention is not particularly limited as long as beingpharmacologically acceptable, and examples of the salt include, forexample, an alkali metal salt such as a sodium salt, and a potassiumsalt; an alkaline earth metal salt such as a calcium salt, and amagnesium salt; an organic base salt such as an ammonium salt, and atrialkylamine salt; a mineral acid salt such as a hydrochloride, and asulfate; and an organic acid salt such as an acetate.

Examples of the solvate of the compound represented by the generalformula (1) or the salt thereof of the present invention include ahydrate, and an alcohol solvate (for example, an ethanol solvate).

The compound represented by the general formula (1) of the presentinvention has an asymmetric carbon atom, and therefore has opticalisomers of a R-isomer and a S-isomer, and all the optical isomers areincluded in the present invention.

Ezetimibe (chemical name:(3R,4S)-1-(4-fluorophenyl)-3-{(3S)-3-(4-fluorophenyl)-3-hydroxypropyl}-4-(4-hydroxyphenyl)azetidin-2-one(IUPAC)) can be produced by a method described in J. Org. Chem., 64(10), 3714-3718 (1999), Tetrahedron Lett., 44(4), 801-804 (2003), andthe like. Further, a pharmaceutical product available on the market maybe used.

As indicated in Examples below, when a compound represented by thegeneral formula (1) is used in combination with a cholesterol absorptioninhibitor, each plasma lipid parameter is improved and particularly astrong HDL-C increasing effect is exerted in an evaluation system usinga rat loaded with high cholesterol diet. Therefore, an agent of thepresent invention is useful in the prevention and/or treatment of mixeddyslipidemia, metabolic syndrome, dyslipidemia complicated by diabetes,hypo-HDL cholesterolemia, or hypertriglyceridemia.

The dyslipidemia in the present invention means that any one or two ormore of the total triglyceride (TG) level, the total cholesterol (TC)level, the VLDL cholesterol (VLDL-C) level, the LDL cholesterol (LDL-C)level, or the HDL cholesterol (HDL-C) level, in the blood deviate from arange of the normal level.

In addition, the disease in a subject requiring increase of HDLcholesterol (HDL-C) in the present invention means the case where theHDL-C level in the blood is lower than the normal level.

The compound represented by the general formula (1) or a salt thereof,or a solvate of the compound, or a solvate of the salt of the compoundof the present invention can be prepared alone or in combination withother pharmaceutically acceptable carriers, into a dosage form of atablet, a capsule, granules, powders, lotion, ointment, an injection, asuppository, and the like. These pharmaceutical preparations can beproduced by a known method. For example, a preparation for oraladministration can be produced by the formulation of a solubilizer suchas tragacanth gum, gum arabic, sucrose fatty acid ester, lecithin, oliveoil, soybean oil, and PEG 400; an excipient such as starch, mannitol,and lactose;

-   a binder such as methylcellulose, carboxymethylcellulose sodium, and    hydroxypropylcellulose; a disintegrant such as crystalline    cellulose, and carboxymethylcellulose calcium; a lubricant such as    talc, and magnesium stearate; a fluidity-improving agent such as    light silicic anhydride; and the like, in appropriate combination.

As to the usage form of the pharmaceutical composition of the presentinvention, a) a compound represented by the general formula (1) or asalt thereof, or a solvate of the compound, or a solvate of the salt ofthe compound is used in combination with b) a cholesterol absorptioninhibitor, and the pharmaceutical composition can be used in a form inwhich a prophylactic and/or therapeutic effect for dyslipidemia such asmixed dyslipidemia, metabolic syndrome, dyslipidemia complicated bydiabetes, hypo-HDL cholesterolemia, and hypertriglyceridemia is exertedwith the use of the synergistic blood HDL-C increasing effect by theadministration of both agents in addition to each effect of agents,however, is not limited to these usage forms. The compound representedby the general formula (1) and the cholesterol absorption inhibitor maybe administered at the same time, or may be administered separately atintervals.

The compound represented by the general formula (1) and the cholesterolabsorption inhibitor may be prepared together into a single preparation,or may be prepared separately into each preparation and used as a kit.That is, the pharmaceutical composition of the present invention may bea kit composed of an agent containing as an active component at leastone kind selected from a compound represented by the general formula (1)or a salt thereof, or a solvate of the compound, or a solvate of thesalt of the compound, and an agent containing at least one kind ofcholesterol absorption inhibitors in combination.

When both agents are administered as a single preparation in the presentinvention, the mixing ratio of the compound represented by the generalformula (1) to the cholesterol absorption inhibitor can be appropriatelyselected in the range of effective dose of each active component,however, the mixing ratio is generally preferably in the range of 5:1 to1:30,000 by amount ratio, and more preferably in the range of 5:1 to1:1,000 by amount ratio. In particular, in a case where the cholesterolabsorption inhibitor is an agent such as ezetimibe, which suppresses theabsorption of cholesterol by the inhibition of a cholesterol transporter(NPC1-L1), the mixing ratio is preferably in the range of 5:1 to 1:300,and more preferably 3:1 to 1:200 by amount ratio from the viewpoint thatthe particularly excellent synergistic effect is obtained.

In a case where the compound represented by the general formula (1) andthe cholesterol absorption inhibitor are separately prepared, the dosageforms of both preparations may be the same as or different from eachother. Further, the number of administrations of each component mayvary.

The compound represented by the general formula (1) or a salt thereof,or a solvate of the compound, or a solvate of the salt of the compoundof the present invention is orally or parenterally administered. Thedosage of the pharmaceutical of the present invention varies dependingon the body weight, age, sex, symptoms and the like of a subject,however, usually in a case of an adult human, the dosage of 0.001 to 100mg, and preferably 0.01 to 10 mg is preferably administered while beingdivided into 1 to 3 times per day as for the compound of the generalformula (1). Further, the cholesterol absorption inhibitor is preferablyadministered in a dosage of 0.01 to 1,000 mg, and preferably 1 to 100 mgwhile being divided into 1 to 3 times per day as the ezetimibe.

EXAMPLES

Hereinafter, the present invention will be more specifically explainedby way of Examples, Comparative Example, and Preparation Example,however should not be limited at all by these Examples.

Example 1 Combined Effect of Compound A and Ezetimibe for onHigh-Cholesterol Diet-Fed Rats

1. Procedure

SD rats (6 weeks old, male, CHARLES RIVER LABORATORIES JAPAN, INC.) wereused in the experiments. Blood was collected from the jugular vein ofchow-fed rat, and divided into 4 groups (N=8) base on the levels of TGand TC in the plasma, and the body weight. From the next day, a solidchow containing 2% cholesterol and 0.5% sodium cholate (MF-based,Oriental Yeast Co., Ltd.) was loaded to rats for one week. In parallelto the loading with the high cholesterol diet, a vehicle (an aqueoussolution of 0.5% methylcellulose: MC), or(R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyricacid (compound A) and/or ezetimibe was orally administered once a day.In the afternoon of the last day of the administration, blood wascollected under pentobarbital anesthesia after the fasting period of 4hours, and the HDL-C levels in the plasma were measured. The synergisticeffect was determined by using Bürgi's formula (if the relative level inthe combined group is larger than the value of the product of therelative level of single agent A and the relative level of single agentB, the synergistic effect is positive).

2. Group Configuration

Group 1: Control

Group 2: Compound A 0.1 mg/kg

Group 3: Ezetimibe 0.003 mg/kg

Group 4: Compound A 0.1 mg/kg and ezetimibe 0.003 mg/kg

3. Statistical Analysis and Data Processing Method

The results were shown as the mean±standard deviation. Comparisonbetween the control group and the drug administration group wasperformed using Dunnett's multiple comparison test, and the risk rate ofless than 5% was determined to have a significant difference.

4. Results

The results are shown in FIG. 1 and Table 1. In addition, the averagelevel of each group of HDL-C is shown in Table 1. In the singleadministration group of compound A or ezetimibe, apparent effects werenot recognized for the HDL-C level. However, in the combinedadministration group of 0.1 mg/kg of compound A and 0.003 mg/kg ofezetimibe, significant (*:p<0.05, to the control) and synergistic (therelative level (1.167) in the combined group is larger than the value ofthe product (1.010×1.022=1.032) of the relative level in the singleagent A administration group to the control group and the relative levelin the single agent B administration group to the control group)increase of HDL-C level was recognized (FIG. 1, and Table 1).

TABLE 1 Compound A 0.1 mg/kg + Compound A Ezetimibe Ezetimibe Control0.1 mg/kg 0.003 mg/kg 0.003 mg/kg Ave. 24.1 24.3 24.6 28.1 SD 1.6 3.02.4 2.9 HDL-C Rate — 1.0% 2.2% 16.7% of Change

Example 2 Combined Effect of Compound A and Ezetimibe onHigh-Cholesterol Diet-Fed Rats

1. Procedure

SD rats (6 weeks old, male, CHARLES RIVER LABORATORIES JAPAN, INC.) wereused in the experiments. Blood was collected from the jugular vein ofchow-fed rat, and divided into 6 groups (N=6, 8) base on the levels ofTG and TC in the plasma, and the body weight. From the next day, a solidfood containing 2% cholesterol and 0.5% sodium cholate (MF-based,Oriental Yeast Co., Ltd.) was loaded to rats for one week. In parallelto the loading with the high cholesterol diet, a vehicle (0.5% MC), orcompound A and/or ezetimibe was orally administered once a day. In theafternoon of the last day of the administration, blood was collectedunder pentobarbital anesthesia after the fasting period of 4 hours, andthe TG levels and TC levels in the plasma, and lipid levels in thelipoprotein (VLDL-C, LDL-C and HLDL-C) were measured. The synergisticeffect was determined by using Bürgi's formula (in a case where a levellower than that in the control group is indicated in a drugadministration group, that is, when the evaluation of an agent to theTC, VLDL-C, or LDL-C is performed, if the relative level in the combinedgroup is smaller than the value of the product of the relative level ofsingle agent A and the relative level of single agent B, the synergisticeffect exists. In a case where a level higher than that in the controlgroup is indicated in a drug administration group, that is, when theevaluation of an agent to the HDL-C is performed, if the relative levelin the combined group is larger than the value of the product of therelative level of single agent A and the relative level of single agentB, the synergistic effect exists).

2. Group Configuration

Group 1: Control

Group 2: Compound A 0.1 mg/kg

Group 3: Ezetimibe 0.01 mg/kg

Group 4: Combination of compound A 0.1 mg/kg and ezetimibe 0.01 mg/kg

Group 5: Ezetimibe 0.1 mg/kg Group 6: Combination of compound A 0.1mg/kg and ezetimibe 0.1 mg/kg

3. Statistical Analysis and Data Processing Method

The results were shown as the mean±standard deviation. Comparisonbetween the control group and the drug administration group wasperformed using Dunnett's multiple comparison test, and the risk rate ofless than 5% was determined to have a significant difference.

4. Results

The results are shown in Table 2 and FIGS. 2 to 5. FIG. 2 shows theresults of the total cholesterol (TC) levels, FIG. 3 shows the VLDLcholesterol (VLDL-C) levels, FIG. 4 shows the LDL cholesterol (LDL-C)levels, and FIG. 5 shows the HDL cholesterol (HDL-C) levels. The symbolin FIGS. means as follows: *:p<0.05, **:p<0.01, and ***:p<0.001.

Synergistic effects by the combination of compound A with ezetimibe onthe high-cholesterol diet-fed rats

TABLE 2 Compound A (mg/kg) 0.1 — 0.1 — 0.1 Ezetimibe (mg/kg) — 0.01 0.010.1 0.1 TC Relative Level 0.956 0.898 0.635 0.555 0.462 (to Control)Bürgi's Formula 0.859 0.530 VLDL-C Relative Level 0.827 0.719 0.3740.262 0.179 (to Control) Bürgi's Formula 0.595 0.217 LDL-C RelativeLevel 0.971 0.955 0.713 0.720 0.529 (to Control) Bürgi's Formula 0.9270.698 HDL-C Relative Level 1.021 0.910 1.204 1.100 1.326 (to Control)Bürgi's Formula 0.929 1.123

As to each numerical value, the value in the upper shows a relativelevel to the control group, and the value in the lower shows the productof the relative levels in the compound A single administration group andthe ezetimibe single administration group by using Bürgi's formula. Thevalues in the underlined part show that the synergistic effect isrecognized.

Firstly, when the attention was focused on TC (Table 2 and FIG. 2), therelative level of the compound A single administration group was 0.956,the relative level of the 0.01 mg/kg single administration group ofezetimibe was 0.898, and the product of both relative levels was 0.859.On the other hand, the relative level of the combined administrationgroup of compound A and 0.01 mg/kg of ezetimibe was 0.635, and wassignificantly decreased as compared with the product of levels in thesingle administration groups, therefore, the synergistic TC loweringeffect was confirmed.

Further, the relative level of 0.1 mg/kg single administration group ofezetimibe was 0.555, and the product of the relative level 0.555 and therelative level 0.956 of the compound A administration group was 0.530,whereas the relative level of 0.1 mg/kg combined administration group ofthe compound A and the ezetimibe was 0.462, therefore, it was revealedthat the synergistic TC lowering effect was recognized also in thehigh-dose group of ezetimibe.

In the same manner, by the combined administration of the compound A andthe ezetimibe, the synergistic lowering effect for the VLDL-C level andLDL-C level, and the synergistic increasing effect for the HDL-C levelwere recognized.

Comparative Example Combined Effect of Fenofibrate and Ezetimibe onHigh-Cholesterol Diet-Fed Rats

1. Procedure

SD rats (6 weeks old, male, CHARLES RIVER LABORATORIES JAPAN, INC.) wereused in the experiments. Blood was collected from the jugular vein ofchow-fed rat, and divided into 4 groups (N=8) base on the levels of TGand TC in the plasma, and the body weight. From the next day, a solidchow containing 2% cholesterol and 0.5% sodium cholate (MF-based,Oriental Yeast Co., Ltd.) was loaded to rats for one week. In parallelto the loading with the high cholesterol diet, a vehicle (0.5% MC), orfenofibrate (FF) and/or ezetimibe (Ezeti) was orally administered once aday. In the afternoon of the last day of the administration, blood wascollected under pentobarbital anesthesia after the fasting period of 4hours. The HDL-C levels in the plasma were measured.

2. Group Configuration

Group 1: Control

Group 2: Fenofibrate 10 mg/kg

Group 3: Ezetimibe 0.1 mg/kg

Group 4: Combination of fenofibrate 10 mg/kg and ezetimibe 0.1 mg/kg

3. Statistical Analysis and Data Processing Method

The results were shown as the average level±standard deviation.Comparison between the control group and the drug administration groupwas performed using Dunnett's multiple comparison test, and the riskrate of less than 5% was determined to have a significant difference.

4. Results

The results are shown in FIG. 6. Fenofibrate has also been known as aPPARα agonist, however, the synergistic increasing effect for the HDL-Clevel was not recognized in the combination of fenofibrate withezetimibe.

Preparation Example Tablets for Administration Containing Compound A andEzetimibe

The tablets for administration, in which compound A and ezetimibe arecontained, can be produced as in the following.

TABLE 3 Component Dosage Compound A 0.05 mg Ezetimibe 10 mgMicrocrystalline Cellulose 37.25 mg Modified food Grade Cornstarch 37.25mg Magnesium Stearate 0.5 mg

All of the compound A and ezetimibe (amount ratio 1:200) and part of thecellulose and cornstarch were mixed, and the mixture was granulated, asa result, 10% cornstarch paste was obtained. The obtained granules weresieved and dried, and the dried granules were mixed with the remainingpart of the cornstarch, and the magnesium stearate. Next, the obtainedgranules were compressed into a tablet containing 0.05 mg of compound Aand 10 mg of ezetimibe per tablet.

INDUSTRIAL APPLICABILITY

The agent and pharmaceutical composition of the present invention havean excellent blood HDL cholesterol increasing effect, and are useful inthe prevention and/or treatment of dyslipidemia, particularly mixeddyslipidemia, metabolic syndrome, dyslipidemia complicated by diabetes,hypo-HDL cholesterolemia, or hypertriglyceridemia, therefore haveindustrial applicability.

The invention claimed is:
 1. A method of treating dyslipidemia in a human patient in need thereof comprising administering to said patient therapeutically effective amounts of (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or a pharmaceutically acceptable salt thereof and ezetimibe, wherein said (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or said pharmaceutically acceptable salt thereof and said ezetimibe are administered to the human patient at the same time, wherein said patient is suffering from a disease selected from the group consisting of, metabolic syndrome, diabetes, hypo-HDL cholesterolemia, and hypertriglyceridemia.
 2. The method according to claim 1, wherein said patient is suffering from metabolic syndrome.
 3. The method according to claim 1, wherein said patient is suffering from diabetes.
 4. The method according to claim 1, wherein said patient is suffering from hypo-HDL cholesterolemia.
 5. The method according to claim 1, wherein said patient is suffering from hypertriglyceridemia.
 6. The method according to claim 1, wherein an amount ratio of (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or a pharmaceutically acceptable salt thereof to ezetimibe is 1:200 to 3:1.
 7. A method of treating dyslipidemia in a human patient in need thereof comprising administering to said patient therapeutically effective amounts of (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or a pharmaceutically acceptable salt thereof and ezetimibe, wherein said (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or said pharmaceutically acceptable salt thereof and said ezetimibe are administered to the human patient at the same time, wherein the human patient is suffering from an elevated total triglyceride level and a low HDL cholesterol level.
 8. A method of increasing HDL-C in a human patient in need thereof comprising administering to said patient therapeutically effective amounts of (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or a pharmaceutically acceptable salt thereof and ezetimibe, wherein said (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or said pharmaceutically acceptable salt thereof and said ezetimibe are administered to the human patient at the same time.
 9. The method according to claim 8, wherein an amount ratio of (R)-2-[3-[[N-(benzoxazol-2-yl)-N-3-(4-methoxyphenoxy)propyl]aminomethyl]phenoxy]butyric acid or a pharmaceutically acceptable salt thereof to ezetimibe is 1:200 to 3:1. 